Inhibition Of Rat Paw Edema Research Articles

Synthesis and biological evaluation of some novel benzoxazin-4-one and quinazolin-4-one derivatives based on anti-inflammatory commercial drugs

Benzoxazine and quinazoline are nitrogen-containing heterocyclic scaffolds found in various biologically active compounds. Due to their diverse biological actions, these heterocyclic rings serve as crucial frameworks for designing medicinal compounds. This study aimed to synthesize and assess in vivo anti-inflammatory, analgesic, and low ulcerogenic potential of a few novel benz[d][1,3]-oxazin-4-one and quinazolinone derivatives. Benzoxazinones (3a-e) were synthesized by cyclizing the carboxylic group (-COOH) of five nonsteroidal anti-inflammatory drugs viz., aceclofenac, ibuprofen, diclofenac, mefenamic acid and ketoprofen (2a-e) with anthranilic acid (1) using dry phosphorus oxychloride (POCl3) in pyridine. The corresponding quinazolinone derivatives (5a-e) were obtained by reacting 3a-e with isonicotinic acid hydrazide (4). Both sets of compounds were evaluated for their anti-inflammatory, analgesic effects, and ulcerogenicity in animal models. Structural characterization was performed using spectral analysis. Among the benzoxazinone derivatives, compound 2-(2-((2,6-dichlorophenyl) amino) benzyl)-4H-benzo[d][1,3]oxazin-4-one (3d) exhibited significant anti-inflammatory activity (62.61% inhibition of rat paw edema) and analgesic activity (62.36% protection in acetic acid-induced writhings) with tolerable gastrointestinal toxicity (2.67 ulcerogenicity index) compared to quinazolinone derivatives. The results of anti-inflammatory and analgesic activities of both the series are comparable with the respective, positive control. Compound 3d, a benzoxazinone-diclofenac hybrid, emerged as a lead molecule with potent anti-inflammatory, analgesic activities and moderate gastric toxicity showcasing the promising potential for further development.

Evaluation of the Anti-Inflammatory and Antioxidant, Activities of Parinari Curatellifolia Planch. Ex Benth (Chrysobalanaceae) Methanol Stem Bark Extract

It is used as human food and beverage, and in traditional medicine it is used to treat pneumonia and fever. The aim of this was to investigate its anti-inflammatory and antioxidant activities of Parinari curatellifolia methanol stem bark extract. A 2kg of the dried and powdered plant material was extracted with 80% methanol through cold maceration to obtain the crude methanol extract. The crude methanol extract was sequentially fractionated using n-hexane, ethyl acetate and methanol to yield n-hexane, ethyl acetate and methanol fractions respectively. Acute toxicity study was conducted following the OECD guidelines. The anti-inflammatory activity of the fractions was evaluated was evaluated using inhibition of albumin denaturation, inhibition of egg albumin-induced paw edema in rats and phospholipase A2 (PLA2) inhibition assay models. In vitro antioxidant activity was evaluated using DPPH de-colourizing, Nitric oxide scavenging activity and Ferric Reducing Antioxidant Power (FRAP) assays. Oral acute administration of the extract revealed an LD50 dose > 5000 mg/kg in rats. The extract caused a dose- and time-dependent inhibition of rat paw edema, and a concentration-dependent inhibition of Phospholipase A2 activity. IC50 of the extract for Albumin Denaturation inhibition was 37.22 ug/ml. In the antioxidant assays the IC50 of the extract against DPPH was 25.91 ug/ml, while the Nitric oxide Equivalence (mM)/mg was 42.38.

Anti-Inflammatory Potential of Brucea javanica Fruit

Objective: Inflammation plays a crucial role in the progression of many diseases, excessive inflammation increases immune cell activation, which can destroy tissues and body health. When inflammation occurs, multiple pro-inflammatory mediators are overproduced, leading to a variety of diseases such as rheumatism, diabetes, and cardiovascular problems. Brucea javanica has been used as traditional medicine as a therapy for anti-tumor, amebic dysentery, diarrhea, malaria, and intestinal inflammation. Therefore, in this study, we explored the effect of ethanol extract Brucea javanica Fruit on anti-inflammation activity in carrageenan-induced rat paw edema and the COX-2 inhibitory effect in vivo.
 Methods: In vivo anti-inflammatory activity of the ethanolic extracts was evaluated using the carrageenan-induced rat paws edema method at doses of 50, 100, and 200mg/kg and further investigated the effect of Cyclooxygenase-2 inhibitory using ELISA readers.
 Results: The ethanolic extract of Brucea javanica Fruit exhibited anti-inflammatory activity by oral intake of 50 mg/kg of the ethanolic extract and Brucea javanica inhibited rat paw edema by 50.91% significantly compared to Celecoxib by 58.52%. Moreover, Brucea javanica showed COX-2 inhibition by 16,40% compared to Celecoxib by 20,50%.
 Conclusions: These findings indicate that Brucae javanica Fruit extracts have promising anti-inflammatory activity directed against COX-2 enzymatic activity.
 Keywords: Anti-inflammatory, Brucea javanica, Malur, Buah Makasar, Cyclooxygenase.

Formulation of Topical Flurbiprofen Solid Lipid Nanoparticle Gel Formulation Using Hot Melt Extrusion Technique.

Hot melt extrusion (HME) has been used for the formulation of topical solid lipid nanoparticle (SLN) gel without using any other size reduction technique including high pressure homogenization or sonication. SLN formulation solely using HME has not been applied to other drugs except IBU. Therefore, the purpose of the present study was to formulate FLB SLN solely using HME technique and evaluate the SLN formulation in inflammation animal model. Stable 0.5%w/v FLB SLN gel with particle size < 250nm, PI < 0.3 and EE of > 98% was prepared. Differential scanning calorimetry (DSC) thermogram showed that the drug was converted to amorphous form in the HME process. Additionally, rheological studies demonstrated that FLB SLN gel and marketed FLB gel showed shear thinning property. FLB SLN formulation showed significantly (p < 0.05) higher peak force required to spread the formulation as compared to marketed FLB formulation. Stability studies showed that FLB SLN gel was stable for a month at room temperature and 2-4°C. Moreover, in vitro permeation test (IVPT) and ex vivo skin deposition study results revealed that FLB SLN gel showed significant (p < 0.05) increase in drug deposition in dermal layer and drug permeation as compared to control marketed formulation. Further, in vivo anti-inflammatory study showed equivalent inhibition of rat paw edema using 0.5% w/v FLB SLN gel which has 10 times less strength compared to control formulation. Overall, FLB SLN formulation was successfully manufactured solely using HME technique which resulted in enhanced the skin permeation of FLB and superior anti-inflammatory activity.

The essential oil of Melaleuca leucadendra L. (Myrtaceae) from Fatick (Senegal) as a source of methyleugenol. Composition, antibacterial and anti-inflammatory activities

ABSTRACT This study aimed to determine the chemical composition, antibacterial and anti-inflammatory activities of Melaleuca leucadendra leaf essential oils. Ten samples of leaves from M. leucadendra were collected on the salt zone of Fatick (Senegal). The corresponding oils were obtained by hydrodistillation and analyzed by GC/FID and GC/MS. Oil yields from dried leaves were ranged from 1.64% to 2.75%. The almost exclusive compound in all samples of M. leucadendra oils was methyleugenol (98.4–99.5%). The oil exhibited moderate activity against S. aureus and E. coli and no activity against E. faecalis and P. aeruginosa. Anti-inflammatory properties of oil (25, 100 and 150 mg/kg, bw) showed significant inhibition of rat paw oedema induced by carrageenan at the 3rd and 5th h (p < 0.001 and 0.0001, respectively) when compared with control and standard drug (acetylsalicylic acid). The experimental study revealed that M. leucadendra oil, a natural source of methyleugenol, displayed remarkable anti-inflammatory activity.

ANALGESIC AND ANTI-INFLAMMATORY ACTIVITIES OF ETHANOLIC EXTRACT OF LEAVES OF PUNICA GRANATUM L. ON EXPERIMENTAL ANIMAL MODELS

Objective: The aim of the study is to evaluate the analgesic and anti-inflammatory activities of the ethanolic extract of Punica granatum L. (EEPG) on experimental animal models. Methods: Tail-flick method was used to test the central analgesic activity, using Pethidine as standard drug. The tail flick latencies or the basal reaction time of the animals were assessed using an analgesiometer. Glacial acetic acid induced writhing response was used to test the peripheral analgesic activity, using Aspirin as standard drug. Number of writhing responses was counted for 20 min in each group and the percentage protection was calculated. And Carrageenan induced rat paw edema method was used to test anti-inflammatory activity of EEPG against acute inflammation, using Aspirin as standard drug. The inhibition of rat paw edema was calculated in percentage. Results: In central analgesic activity, the extract and pethidine showed significant increase in the reaction time. In peripheral analgesic activity, the extract and aspirin significantly reduced the number of writhes induced by acetic acid. And in anti-inflammatory activity, the extract produced significant reduction of the carrageenan induced paw edema. Conclusion: The EEPG has demonstrated significant analgesic and anti-inflammatory activity.

Ethosomes-based gel formulation of karanjin for treatment of acne vulgaris: in vitro investigations and preclinical assessment.

The online version contains supplementary material available at 10.1007/s13205-021-02978-3.

Anti-inflammatory effect of leaves of Vernonia zeylanica in lipopolysaccharide-stimulated RAW 264.7 macrophages and carrageenan-induced rat paw-edema model

Ethnopharmacological relevanceVernonia zeylanica (L.) Less (Family: Compositae) is a medicinal plant used as external applications for boils, bone fractures, eczema and internally for asthma in traditional medicine in Sri Lanka. Anti-nociceptive, anti-bacterial and anti-proliferative activities have been reported previously. Aim of the studyTo investigate the anti-inflammatory activity of methanol/dichloromethane extract (MDE) of leaves of V. zeylanica by assessing in vivo inhibition of rat paw-edema, in vitro inhibition of the production of nitric oxide (NO) and superoxide and inhibitory effect on inducible nitric oxide synthase (iNOS) gene expression. Materials and methodsIn vivo anti-inflammatory activity of MDE was tested at the dose of 1500 mg/kg using rat paw-edema model. Indomethacin and Gum acacia was used as the positive and vehicle control respectively. In vitro NO inhibitory activity of 7.8–250 μg/ml MDE was tested using lipopolysaccharide (LPS)-stimulated (1 μg/ml) mouse macrophages (RAW264.7 cells) and rat peritoneal cells (RPC) obtained following carrageenan-induction (5 mg/Kg). Griess method was used to quantify the nitrite levels in culture supernatants. In vitro inhibition of superoxide production of Phorbal 12-myristate 13-acetate (PMA)-stimulated RAW cells was determined by quantitative Nitroblue Tetrazolium (NBT) assay. N-monomethyl-L-arginine acetate (NMMA) (1 mM) and Diphenyleneiodonium chloride (DPI) (10 μM) were used as the positive controls for inhibitory activity of NO and superoxide production respectively. Reverse transcriptase polymerase chain reaction (RT-PCR) analysis was carried out to test the inhibitory effect on mRNA expression of iNOS gene. ResultsTreatment with MDE of V. zeylanica at 1500 mg/kg showed significant inhibition of paw-edema from 1st-5th hour (P < 0.01) compared with the control. The reference drug, indomethacin showed a biphasic pattern and its highest inhibition was (98.3 ± 7.1%) at 4th h (P < 0.01). MDE of V. zeylanica showed similar inhibition of paw-edema with highest inhibition recorded as 94.5 ± 5.28%, at 5th h (P < 0.01). The inhibitory concentration (IC50) of MDE for in vitro NO inhibitory activity was 105 μg/ml for RAW cells and 80 μg/ml for RPCs. Both NO inhibitory activities showed significant dose-dependency (r = 0.998 and r = 0.915 respectively; p < 0.05). MDE concentration of 250 μg/ml showed 55% inhibition of ROS production in RAW cells. NMMA showed 78% and 70.1% inhibition of NO production with RAW cells and RPCs whereas DPI showed 61% superoxide inhibitory activity with RAW cells. NO inhibitory activity of MDE on RAW cells was confirmed by the significant reduction (99.1%) in iNOS gene expression. ConclusionThese results demonstrated potent anti-inflammatory activity of MDE of V. zeylanica reflected by its significant in vivo inhibition of rat paw-edema, in vitro inhibition of NO and superoxide production, and the reduction of iNOS gene expression. Thus, further purification and isolation of bioactive compounds from V. zeylanica are emphasized.

In-vivo Anti-inflammatory testing of extracts and its fractions obtained from Saptaparn leaves (Alstonia scholaris Linn. R. Br)

Since antiquity, Alstonia scholaris commonly known as devil's tree or blackboard tree has been used for the treatment of many human ailments. The plant is used in Ayurvedic, Unani and Sidhha/Tamil types of alternative medicinal systems. Literature suggests that Alstonia scholaris is useful in treating malaria, abdominal disorders, dyspepsia, leprosy, skin diseases, tumors, chronic and foul ulcers, asthma, bronchitis etc. In order to validate these ethno-botanical practices the anti-inflammatory activity of methanolic extracts and various bioactive guided fractions of leaf of Saptparn (Alstonia scholaris, Apocyanaceae) tested. The anti-inflammatory activity of the extracts and its various fractions were evaluated using the carrageenan-induced rat paw edema method for the determination of acute phase of inflammation. Indomethacin used as reference drug. The result of pharmacological studies (anti-inflammatory) indicated the methanolic extract (200 mg/kg) significantly inhibited carrageenan induced rat paw edema in comparison to the inhibition of edema by standard drug Indomethacin (10mg/kg). Maximum inhibition of rat paw edema was observed with methanolic extract of plant (400 mg/kg) at the end of 4th hour when compared to control group. The chloroform fraction (50 mg/kg and 100 mg/kg) possess statistically significant anti-inflammatory activity. Maximum inhibition of rat paw edema was observed with chloroform fraction at 100 mg/kg at the end of 5th hour when compared to control group.

Design and synthesis of sulfur cross-linked 1,3,4-oxadiazole-nitro(furan/thiophene)-propenones as dual inhibitors of inflammation and tuberculosis: molecular docking and Hirshfeld surface analysis

A series of 3-[5-nitro(furan/thiophene)-2-yl]-1-aryl-3-(5-aryl-1,3,4-oxadiazol-2-ylthio)prop-2-en-1-one derivatives was synthesized and studied with the aim of developing dual inhibitors of multidrug-resistant tuberculosis and inflammation. The in vivo anti-inflammatory activity results showed excellent inhibition of rat paw edema. The methoxybenzene/nitrofuryl derivative of title compounds showed 83% inhibition of inflammation during 2–6 h after carrageenan injection. All compounds showed anti-tuberculosis activity at MIC of 50 µg/cm3. The molecular docking studies revealed that the oxadiazole and nitrofuran groups played a significant role in the inhibiting site of the enzymes COX1, COX2, 5-LOs, and InhA by forming hydrogen bonding with Tyr 385, Ser 530, Tyr 467, and Tyr 158 amino acid residues, respectively. The novel compounds are active antibacterial agents with potential inhibition on E. coli bacteria. The toxicity results showed good percentage viability of human kidney cell lines with IC50 value greater than 100 µg/cm3 concentration. The Hirshfeld surface analysis and electrostatic potential map of compound showed good intermolecular contacts and hydrogen bonding donor and acceptor potential.

Enhanced transdermal delivery of lornoxicam by nanostructured lipid carrier gels modified with polyarginine peptide for treatment of carrageenan-induced rat paw edema.

Background: Nanostructured lipid carriers (NLCs) are emerging as attractive drug carriers in transdermal drug delivery. The surface modification of NLCs with cell-penetrating peptides (CPPs) can enhance the skin permeation of drugs.Purpose: The objective of the current study was to evaluate the ability of the cell-penetrating peptide (CPP) polyarginine to translocate NLCs loaded with lornoxicam (LN) into the skin layers and to evaluate its anti-inflammatory effect.Methods: The NLCs were prepared using an emulsion evaporation and low temperature solidification technique using glyceryl monostearates, triglycerides, DOGS-NTA-Ni lipids and surfactants, and then six histidine-tagged polyarginine containing 11 arginine (R11) peptides was modified on the surface of NLCs.Results: The developed NLCs formulated with LN and R11 (LN-NLC-R11) were incorporated into 2% HPMC gels. NLCs were prepared with a particle size of (121.81±3.61)–(145.72±4.78) nm, and the zeta potential decreased from (−30.30±2.07) to (−14.66±0.74) mV after the modification of R11 peptides. The encapsulation efficiency and drug loading were (74.61±1.13) % and (7.92±0.33) %, respectively, regardless of the surface modification. Cellular uptake assays using HaCaT cells suggested that the NLC modified with R11 (0.02%, w/w) significantly enhanced the cell internalization of nanoparticles relative to unmodified NLCs (P<0.05 or P<0.01). An in vitro skin permeation study showed better permeation-enhancing ability of R11 (0.02%, w/w) than that of other content (0.01% or 0.04%). In carrageenan-induced rat paw edema models, LN-NLC-R11 gels inhibited rat paw edema and the production of inflammatory cytokines compared with LN-NLC gels and LN gels (P<0.01).Conclusion: In our investigation, it was strongly demonstrated that the surface modification of NLC with R11 enhanced the translocation of LN across the skin, thereby alleviating inflammation.

Antiinflammatory Activity of Wheat Grass Fortified with Cow Urine Distillate

Cow urine distillate appear to possess bioenhancement capacity that increases effectiveness of medicinal products. So, the current research was aimed to evaluate the anti-inflammatory and antioxidant activity of wheat grass powder fortified with cow urine distillate. Antiinflammatory activity was studied using carrageenan-induced paw edema in Wistar rats at 100, 200 and 400 mg/kg doses of wheat grass powder fortified with cow urine distillate and the antioxidant capacity was studied using the ferric thiocyanate method. Wheat grass powder fortified with cow urine distillate showed significant reduction (61.09 %) in peroxide production in the ferric thiocyanate method. All the three doses tested produced significant inhibition of rat paw edema in the carrageenan assay but the 400 mg/kg dose produced maximum inhibition of rat paw edema (40.06 %) at the 5 h time point compared to the control. It is possible to conclude from the results of the study concludes that fortification of wheat grass powder with cow urine distillate enhanced the antioxidant and antiinflammatory activities and further research is necessary to understand the mechanism and phytochemicals responsible for the fortification of the activity of wheat grass.

Optimization of ethosomes for topical thymoquinone delivery for the treatment of skin acne

The current study involved the development and optimization of ethosomes formulation for topical delivery of thymoquinone for improved therapy in skin acne. Formulation was optimized based on vesicles size, entrapment efficiency and flux by applying “3-factor 3-level Box-Behnken design”. The optimized formulation was further evaluated for “anti-microbial, anti-inflammatory, anti-acne and skin irritation activity”. The optimized formulation had shown the vesicles size of 105.2 ± 8.0 nm, entrapment efficiency of 85.30 ± 6.30% and flux of 65.40 ± 7.6 μg/cm2/h. The penetration depth of rhodamine loaded ethosomes across rat skin was traced with confocal laser microscopy and it was noted that the plain rhodamine solution (control) remained confined to few micrometers (14.4 μm) depth only into rat skin, while the rhodamine loaded ethosomes formulation penetrate deeper up to 103.5 μm depth into rat skin. Antimicrobial study exhibited that the marketed allopathic gel, prepared ethosomes formulation, and marketed herbal gel have presented zone of inhibition of 29.0 ± 1.00 mm, 20.33 ± 0.57 mm, and 15.00 ± 1.73 mm respectively against S. Epidermidis. The in vivo anti-inflammatory activity demonstrated that the application of prepared ethosomes and marketed diclofenac gel formulation resulted in an inhibition of rat paw edema by 80.74% and 84.61% respectively in Wistar rats. Anti-acne study demonstrated that the developed ethosomes formulation had shown considerable effect on sebaceous glands units by reducing its glands number and size. Further skin irritation study revealed that the developed formulation was safe, less irritant and well tolerable formulation for topical delivery. It was concluded that the developed thymoquinone loaded ethosomes formulation can be used as an effective treatment option for acne vulgaris and various skin disorders.

Preparation and pre-clinical characterization of sustainedrelease ketoprofen implants for the management of pain and inflammation in osteoarthritis

Purpose: To prepare and evaluate sustained-release ketoprofen implants for prolonged drug release and activity.Methods: Ketoprofen implants were prepared with poly (lactic-co-glycolic acid) (PLGA) and chitosan in the form of tablets. The implants were analyzed for drug loading, thickness, hardness, swelling, in vitro drug release, as well as in vivo analgesic and anti-inflammatory activities.Results: The implants were round, smooth in appearance, uniform in thickness and showed no cracks or physical defects on the surface. Their friability was &lt; 1 % while drug content ranged from 89.98 ± 2.06 to 92.95 ± 1.65 %. In vitro drug release ranged from 70.23 to 92.04 % at the end of 5 days. Implants containing higher amounts of PLGA produced the highest swelling (40.24 ± 1.08 %). Implant IKT3 showed maximum analgesic activity (7.75 ± 1.00 s) and shortest time of maximum analgesia (2.5 h) in hot plate method. Inhibition of rat paw edema for IKT1, IKT2 and IKT3 was 79.95, 69.98 and 82.24 %, respectively, after 24 h.Conclusion: Ketoprofen-loaded implant IKT3 (4:4:2 ratio of PLGA, chitosan and ketoprofen) provides relatively quick onset and prolonged duration of analgesic effect. Thus, ketoprofen implants have a potential for development into therapeutic products for prolonged management of pain and inflammation in osteoarthritis.Keywords: Osteoarthritis, Ketoprofen implant, Prolonged analgesia, Poly(lactic-co-glycolic acid), Chitosan

SYNTHESIS, ANTIOXIDANT AND ANTI-INFLAMMATORY ACTIVITIES OF ETHYL 2-(2-CYANO-3- (SUBSTITUTED PHENYL)ACRYLAMIDO)-4,5-DIMETHYLTHIOPHENE-3-CARBOXYLATES

Objective: Objective of the study was to synthesize and evaluate a series of novel compounds, ethyl 2-(2-cyano-3-(substituted phenyl)acrylamido)- 4,5-dimethylthiophene-3-carboxylates, for in vitro antioxidant and in vivo anti-inflammatory activities.Methods: Ethyl 2-(2-cyano-3-(substituted phenyl)acrylamido)-4,5-dimethylthiophene-3-carboxylates were synthesized by knoevenagel condensation of active methylene group of ethyl 2-(2-cyanoacetamido)-4,5-dimethylthiophene-3-carboxylate with substituted benzaldehydes. The synthesized compounds were evaluated for their in vitro antioxidant properties in three different models, viz., reduction of 1,1-diphenyl-2-pycrylhydrazyl free radical, scavenging of nitric oxide free radical, and ferric ion-induced lipid peroxidation using rat brain homogenate. Few selected compounds with good antioxidant properties were pharmacologically evaluated for anti-inflammatory activity by carrageenan-induced rat paw edema model.Results: Clean and efficient synthetic procedure was used for the preparation of series of compounds. The structures of synthesized compounds were confirmed by infrared, 1H nuclear magnetic resonance and mass spectra. The antioxidant activity data revealed that the compounds of ethyl 2-(2-cyano-3-(substituted phenyl)acrylamido)-4,5-dimethylthiophene-3-carboxylate containing phenolic substitution showed greater antioxidant activity. Hence, the active compounds were evaluated for anti-inflammatory activity and found to possess good activity. The percentage inhibition of rat paw edema obtained for the evaluated compounds was in the range of 70.2-83.1, comparable to the standard drug diclofenac (85.0%).Conclusion: The use of inexpensive, eco-friendly and readily available reagents, easy work-up and high purity of products makes the procedure a convenient and robust method for the synthesis of title compounds. The compounds of ethyl 2-(2-cyano-3-(substituted phenyl)acrylamido)-4,5- dimethylthiophene-3-carboxylate containing phenolic substitution showed greater antioxidant and anti-inflammatory activities.

Nanoparticle formulation of 11-keto-β-boswellic acid (KBA): anti-inflammatory activity and in vivo pharmacokinetics

Context: The oleo-gum-resin of Boswellia serrata Roxb. (Burseraceae) is widely used for the treatment of inflammatory diseases such as osteoarthritis, rheumatoid arthritis and cancer. Anti-inflammatory activity of 11-keto-β-boswellic acid (KBA) is impeded by poor oral bioavailability due to its high lipid solubility, rapid phase-1 metabolism and poor intestinal permeability.Objective: This study developed a poly-dl-lactide-co-glycolide-based nanoparticle formulation of KBA to improve its oral bioavailability and in vivo anti-inflammatory activity.Materials and methods: KBA was isolated from the oleo-gum resin of B. serrata, and its nanoparticle formulation (KBA-NPs) was prepared by the emulsion–diffusion–evaporation method. Oral bioavailability of KBA and KBA-NPs was studied at 50 mg/kg p.o. dose in Sprague–Dawley rats, and further evaluated for in vivo anti-inflammatory activity in carrageenan-induced rat paw oedema assay at the same dose level.Results: The prepared KBA-NPs had a particle size of 152.6 nm with polydispersity index of 0.194, 79.7% entrapment efficiency and a cumulative 61.5% release of KBA from KBA-NPs, at 72 h. KBA-NPs showed 60.8% inhibition of rat paw oedema at 5 h as compared to 34.9% as that of KBA. The results of oral bioavailability study and in vivo anti-inflammatory activity showed 7- and 1.7-fold increase in bioavailability and anti-inflammatory activity, respectively, of KBA in KBA-NPs as compared to KBA alone.Conclusion: The results of improved oral bioavailability and in vivo anti-inflammatory activity of KBA-NPs suggested successful development of KBA nanoparticle formulation.

Intrathecal Delivery of Ketorolac Loaded &lt;i&gt;In Situ&lt;/i&gt; Gels for Prolonged Analgesic and Anti-Inflammatory Activity in Vertebral Fracture

Purpose: To develop biodegradable, polymeric in situ gels based on sodium alginate and hydroxypropyl methylcellulose for intrathecal delivery of ketorolac tromethamine (KT) for effective management of pain and inflammation in vertebral fracture.Method: Ion activated in situ gels were used as implants and were prepared from sodium alginate and hydroxypropyl methylcellulose. The fabricated gels were evaluated for visual appearance, clarity, pH, gelling capacity, drug content, viscosity (using Brookfield viscometer), in vitro drug release (using a fabricated KC cell) and in vivo analgesic and anti-inflammatory activity (by intrathecal administration of in situ gel near the fractured vertebra in a rat model).Results: The physicochemical properties (visual appearance, clarity, pH, gelling capacity, drug content and viscosity) of in situ gels were acceptable for therapeutic use. KT-loaded gels demonstrated high drug encapsulation efficiency (98.3 - 103.3 %). Further, KT-loaded gels exhibited viscosity in the range of 1.11 to 6 cps at 50 rpm and shear thinning property (rheology testing). Additionally, the gels demonstrated 84.43 to 96.98 % drug release at the end of 12 h. In particular, in situ gels prepared from 1.2 % alginate/0.4 % HPMC (G7) exhibited excellent analgesic (54.28 %) and anti-inflammatory activity (51.6 % inhibition of rat paw edema) in the animal model of vertebral fracture.Conclusion: The formulated in situ gels can potentially be used as implants for the treatment of patients with vertebral fracture.Keywords: Ketorolac, Orthopaedic implant, Extended release, Analgesic, Anti inflammation, Vertebral fracture

Analgesic and anti-inflammatory activity of ?-sitosterol isolated from leaves of Oxalis corniculata

Oxalis corniculata Linn. (Oxalidaceae) is one of the important medicinal plants used traditionally for the treatment of fever, pain and inflammation. To validate folk use of Oxalis corniculata as analgesic and anti-inflammatory remedy. The leaves of Oxalis corniculata was used for successive extraction with increasing polarity solvents. Petroleum ether extract was selected for activity guided fractionation to isolate ?-sitosterol due to its better efficacy than other extracts. Analgesic activity was done by hot plate test and acetic acid-induced writhings, while anti-inflammatory activity by carrageenan-induced paw edema method. All the extracts were screened at the dose of 100 mg/kg, i.p. and isolated ?-sitosterol was screened at the doses of 5, 10 and 20 mg/kg, i.p. Naloxone (1 mg/kg, s.c.) was used to understand the mechanism of nociception. In hot plate test, Petroleum ether extract was found most active with reaction time of 8.4 0.4 sec after 60 min while isolated ?-sitosterol at dose of 20 mg/kg showed 11.1 0.3 sec after 90 min. The number of writings in 30 min was compared with paracetamol. Petroleum ether extract and ?-sitosterol (20 mg/kg) showed 43.14 1.9 and 34.21 1.4 writhings respectively. Isolated ?-sitosterol (20 mg/kg) inhibited rat paw edema to 0.32 0.06 ml after 120 min. Naloxone reversed antinociceptive effects of extracts and isolated ?-sitosterol. It can be concluded that isolated ?-sitosterol is responsible for analgesic and anti-inflammatory activity of Oxalis corniculata leaves and it works through central mechanism.

Penetration Enhancement Effect of Turpentine Oil on Transdermal Film of Ketorolac

Purpose: To prepare transdermal films of ketorolac tromethamine (KT) and study the effect of turpentine oil as a penetration enhancer for the drug. Methods: Transdermal films of KT were prepared with Carbopol-934 and ethyl cellulose, with turpentine oil as the penetration enhancer, using solvent evaporation method. The films were characterized for physicochemical properties, ex vivo permeation, as well as in vivo anti-inflammatory and analgesic activities in Wistar rats. Results: The transdermal films were uniform in weight and thickness, flat, with high drug content (93.9 to 98.5 %) and of high folding endurance (134.0 to 180.0). Drug permeation through excised rat abdominal skin was prolonged, with the total drug release ranging from 58.88 to 88.98 % in 24 h. The films containing penetration enhancer showed higher drug permeation than the one without the enhancer; furthermore, drug permeation increased with increase in the concentration of the enhancer. The films were non-irritant to the skin. The transdermal films prepared with permeation enhancers showed greater anti-inflammatory activity (87.55 ± 2.50 and 83.24 ± 2.29 % inhibition of rat paw edema at the end of 12 h for formulations F2 and F3, respectively, compared to that of the formulation without enhancer with 69.99 %) as well as greater analgesic activity (quicker onset of analgesia in 1.5 h with longer duration of 10 to 12 h). Conclusion: Transdermal films of ketorolac have a potential for use in the treatment of pain andinflammation. Incorporation of turpentine oil in the films enhances not only drug flux but also analgesic and anti-inflammatory activities in rats.

Mechanisms involved in the anti-inflammatory action of a polysulfated fraction from Gracilaria cornea in rats.

The anti-inflammatory mechanisms of the sulfated polysaccharidic fraction obtained from red marine alga Gracilaria cornea (Gc-FI) were investigated using a paw edema model induced in rats by different inflammatory agents (carrageenan, dextran, serotonin, bradykinin, compound 48/80 or L-arginine). Gc-FI at the doses of 3, 9 or 27 mg/kg, subcutaneously - s.c., significantly inhibited rat paw edema induced by carrageenan and dextran, as confirmed by myeloperoxidase and Evans’ blue assessments, respectively. Gc-FI (9 mg/kg, s.c.) inhibited rat paw edema induced by histamine, compound 48/80 and L-arginine. Additionally, Gc-FI (9 mg/kg, s.c.) inhibited Cg-induced edema in animals with intact mast cells but did not inhibit that with degranulated mast cells by compound 48/80, revealing a protective role on mast cell membranes. Gc-FI down-regulated the IL-1β, TNF-α and COX-2 mRNA and protein levels compared with those of the carrageenan group, based on qRT-PCR and immunohistochemistry analyses. After inhibition with ZnPP IX, a specific heme oxygenase-1 (HO-1) inhibitor, the anti-inflammatory effect of Gc-FI was not observed in Cg-induced paw edema, suggesting that the anti-inflammatory effect of Gc-FI is, in part, dependent on the integrity of the HO-1 pathway. Gc-FI can target a combination of multiple points involved in inflammatory phenomena.